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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 楊姍樺 | zh_TW |
| dc.contributor.advisor | Shan-Hua Yang | en |
| dc.contributor.author | 王榆茜 | zh_TW |
| dc.contributor.author | Yu-Chien Wang | en |
| dc.date.accessioned | 2026-02-26T16:54:17Z | - |
| dc.date.available | 2026-02-27 | - |
| dc.date.copyright | 2026-02-26 | - |
| dc.date.issued | 2026 | - |
| dc.date.submitted | 2026-02-02 | - |
| dc.identifier.citation | 劉莉蓮, 蔡鋒樺, 塗子萱, 徐聖翔, 賴威丞, 林恩如, 陳品嶧, & 林儀恩. (2022). 110年度臺灣硨磲貝及其他重要螺貝類調查及保育評估計畫.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/101710 | - |
| dc.description.abstract | 硨磲貝是珊瑚礁生態系中獨特的底棲生物，其殼體可供海洋生物庇護，而排泄物中所含共生藻能被珊瑚或硨磲貝幼體吸收，作為自身共生藻使用。硨磲貝與體內微生物共同構成共生體，其中共生藻和細菌與宿主間具有高度相關性。然而，近年來全球暖化使水溫急遽上升，導致硨磲貝面臨環境壓力。此外，弧菌屬為海洋生物常見致病菌，前人研究顯示，高溫加熱而瀕臨死亡的硨磲貝體內，弧菌屬相對豐度高於健康個體。雖然珊瑚與其體內微生物的相互作用已被廣泛研究，但針對硨磲貝在季節性溫度變化下共生微生物的群落組成與變化仍相對缺乏，而共生藻與細菌在組織中的空間分佈亦尚無更詳盡的描述。因此本研究旨在探討硨磲貝於不同季節下之共生微生物 (共生藻、共棲細菌) 的群落組成，並利用qPCR及FISH定量方法分析弧菌屬豐度變化。同時，透過FISH觀察共生藻與細菌在組織中的空間分佈。此研究為排除個體間之差異，均採用已標定硨磲貝之外套膜進行分析。結果顯示，於春季時，共生藻之群落組成發生改變，且Clade C (Cladocopium)與Clade D (Durusdinium) 相對豐度呈現上升趨勢。在細菌群落方面，弧菌屬相對豐度於夏季時較高，冬季則呈現下降趨勢，而弧菌屬qPCR定量顯示其在秋季時的含量較其他階段高，但未觀察到顯著差異。此外，FISH定量結果與次世代定序具有一致性，且於影像中觀察到細菌與共生藻在外套膜內具有高度緊密之空間分佈關係，暗示共生微生物之間亦具有共存關係，未來研究可深入探討兩者間的交互作用。 | zh_TW |
| dc.description.abstract | Giant clams are special benthic invertebrates in coral reef ecosystem. Their shells can provide refuges for reef organisms, and fecal pellets contain viable Symbiodiniaceae that can be acquired by coral and giant clams’ larvae and maintained as their own symbionts. Giant clams form a holobiont together with their associated microbiome. However, global warming–driven increases in seawater temperature have led to environmental stress in giant clam holobionts. Vibrio are common pathogen of marine organisms, and previous studies have shown that the relative abundance of Vibrio is higher in dying giant clams than in healthy individuals. Although coral-associated microbiome interaction has been widely studied, shift in the composition and abundance of giant clam’s microbiome across seasonal temperature variations remain less explored, particularly regarding the spatial distribution of Symbiodiniaceae and bacteria within host tissues. Therefore, this study aims to investigate the Symbiodiniaceae and bacteria community dynamics of giant clams across different season. Moreover, qPCR and FISH were employed to quantify changes in Vibrio abundance, while FISH imaging was used to examine the tissue-level spatial distribution. To control for individual variation, each giant clam in the field was tagged to ensure repeated sampling of the same individuals. The results demonstrate that Symbiodiniaceae community composition changed in spring, with increased relative abundance of Clade C (Cladocopium) and Clade D (Durusdinium). Relative abundance of Vibrio was higher in summer but decrease in winter; qPCR indicates higher abundance in autumn. FISH result aligned with sequencing and revealed close spacial distribution between Symbiodiniaceae and bacteria. This suggests that symbiotic microbiome may also exhibit a coexistence relationship, and future studies could investigate the interactions between the two. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2026-02-26T16:54:17Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2026-02-26T16:54:17Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 口試委員審定書 I
謝辭 II 摘要 III Abstract IV 目次 V 圖次 XI 表次 XIII 壹、前言 1 1. 硨磲貝及其生態系介紹 1 1.1. 珊瑚礁生態系 1 1.2. 硨磲貝的獨特性 2 1.3. 硨磲貝的基本構造 4 1.4. 硨磲貝的現況 4 2. 硨磲貝共生體 (Giant clam holobiont) 6 2.1. 共生體的概念 6 2.2. 共生藻與宿主的相互作用 7 2.3. 細菌與宿主的相互作用 8 3. 環境溫度變動下硨磲貝共生體的變化 9 3.1. 硨磲貝宿主之生理表現 9 3.2. 硨磲貝共生體之變化 10 4. 研究動機與目的 11 貳、材料與方法 14 1. 樣本採集和固定 14 2. 硨磲貝外套膜共棲微生物群落分析 14 2.1. 去氧核糖核酸 (Deoxyribonucleic acid, DNA) 萃取 14 2.2. 聚合酶連鎖反應 (Polymerase Chian Reaction, PCR) 15 2.2.1. 共棲細菌16S 核醣體RNA基因擴增 15 2.2.2. 共生藻ITS2區域擴增 16 2.3. DNA條碼基因擴增 (DNA Barcoding PCR) 17 2.4. 產物純化及定序文庫製備 18 2.4.1. 共棲菌樣本純化及文庫製備 18 2.4.2. 共生藻樣本純化及文庫製備 18 2.5. 定序資料處理與統計分析 19 2.5.1. α多樣性 (Alpha diversity) 20 2.5.2. β多樣性 (Beta diversity) 20 2.5.3. 相對豐度 (Relative abundance) 21 2.5.4. 季節間獨有及共有ASV分析 21 2.5.5. 菌相潛在功能預測分析 22 3. 螢光原位雜合 (Fluorescent in situ hybridization, FISH) 23 3.1. 石蠟包埋與切片 23 3.2. 樣本脫蠟 24 3.3. 穿透處理 24 3.4. 探針雜合 (Probe hybridization) 25 3.5. DAPI (4', 6-diamidino-2-phenylindole) 染色 26 3.6. 樣本封片 26 3.7. 共軛焦顯微鏡 (Confocal microscope) 觀察 26 3.8. 影像計數分析及統計分析 27 4. 定量聚合酶連鎖反應 (Quantitative Polymerase Chian Reaction, qPCR) 27 4.1. 弧菌屬之定量聚合酶連鎖反應專一性探針 (qPCR probe) 27 4.2. 設計弧菌屬之qPCR引子對 (qPCR primer) 29 4.3. 實驗菌株製備 29 4.3.1. 菌株挑選 29 4.3.2. 菌株活化及培養 30 4.3.3. 菌株之DNA萃取 31 4.4. qPCR條件預試 32 4.4.1. qPCR引子對專一性驗證 32 4.4.2. qPCR探針專一性驗證 33 4.4.3. qPCR最佳黏合溫度篩選 34 4.5. 製備檢量線 35 4.6. 定量聚合酶連鎖反應 35 參、結果 37 1. 野外採樣水下溫度紀錄 37 2. 硨磲貝外套膜共生藻分析 37 2.1. 硨磲貝共生藻α多樣性 (Alpha diversity) 37 2.2. 硨磲貝共生藻β多樣性 (Beta diversity) 38 2.3. 硨磲貝共生藻相對豐度 (Relative abundance) 38 2.3.1. 屬 (Genus) 階層的硨磲貝共生藻相對豐度 38 2.3.2. 亞群 (Subclade) 階層的硨磲貝共生藻相對豐度 39 3. 硨磲貝外套膜之共棲細菌分析 40 3.1. 硨磲貝共棲細菌α多樣性 (Alpha diversity) 40 3.2. 硨磲貝共棲細菌β多樣性 (Beta diversity) 40 3.3. 硨磲貝共棲細菌相對豐度 (Relative abundance) 40 3.3.1. 目 (Order) 階層的硨磲貝共棲細菌之相對豐度 40 3.3.2. 科 (Family) 階層的硨磲貝共棲細菌之相對豐度 41 3.3.3. 屬 (Genus) 階層的硨磲貝共棲細菌之相對豐度 42 3.3.4. 不同季節間具有顯著差異之細菌ASV的相對豐度 43 3.4. 硨磲貝共棲細菌共享與獨有ASV數量分佈 43 3.5. 硨磲貝共棲細菌潛在功能預測 43 4. 水樣菌相分析 44 4.1. 水樣菌相α多樣性 (Alpha diversity) 44 4.2. 水樣菌相β多樣性 (Beta diversity) 45 4.3. 屬 (Genus) 階層的水樣菌相相對豐度 (Relative abundance) 46 5. 螢光原位雜合 (Fluorescent in situ hybridization, FISH) 47 6. 定量聚合酶連鎖反應 (Quantitative Polymerase Chian Reaction, qPCR) 47 肆、討論 49 1. 實驗的限制性 49 2. 不同宿主個體與共生體造成白化延遲的機制 50 2.1. 外型結構 50 2.2. 共生藻分佈位置 51 2.3. 共生藻的存活策略 51 3. 季節間硨磲貝共生體的變動 52 3.1. 共生藻群落的重組 52 3.2. 優勢菌群的變動及其功能 53 3.3. 預測潛在功能之相對貢獻值與季節的關聯性 56 4. 不同季節中水樣共棲細菌的變動及其功能 57 5. 外套膜中共生藻－細菌的空間分佈及相關菌群功能角色 58 伍、結論 60 陸、圖與表 61 柒、參考文獻 113 捌、附錄 128 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 硨磲貝 | - |
| dc.subject | 季節 | - |
| dc.subject | 共棲細菌 | - |
| dc.subject | 共生藻 | - |
| dc.subject | 弧菌屬 | - |
| dc.subject | Giant clam | - |
| dc.subject | Season | - |
| dc.subject | Bacteria | - |
| dc.subject | Symbiodiniaceae | - |
| dc.subject | Vibrio | - |
| dc.title | 不同季節硨磲貝共生微生物群落與弧菌屬菌群之動態變化 | zh_TW |
| dc.title | Seasonal Dynamics of Symbiotic Microbial Communities and Vibrio in Giant Clams | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 114-1 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.coadvisor | 蕭仁傑 | zh_TW |
| dc.contributor.coadvisor | Jen-Chieh Shiao | en |
| dc.contributor.oralexamcommittee | 塗子萱;何攖寧 | zh_TW |
| dc.contributor.oralexamcommittee | Tzu-Hsuan Tu;Ying-Ning Ho | en |
| dc.subject.keyword | 硨磲貝,季節共棲細菌共生藻弧菌屬 | zh_TW |
| dc.subject.keyword | Giant clam,SeasonBacteriaSymbiodiniaceaeVibrio | en |
| dc.relation.page | 139 | - |
| dc.identifier.doi | 10.6342/NTU202600504 | - |
| dc.rights.note | 同意授權(全球公開) | - |
| dc.date.accepted | 2026-02-04 | - |
| dc.contributor.author-college | 理學院 | - |
| dc.contributor.author-dept | 海洋研究所 | - |
| dc.date.embargo-lift | 2028-01-30 | - |
| Appears in Collections: | 海洋研究所 | |
Files in This Item:
| File | Size | Format | |
|---|---|---|---|
| ntu-114-1.pdf Until 2028-01-30 | 163.55 MB | Adobe PDF |
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